(A) Recovery of iCM beating rate after cell exposure to UW with ASC-S generated by either control ASC (ASCC), or ASC transfected with silencing RNA directed to SOD1 (ASCSOD1-), SOD3 (ASCSOD3-), catalase (ASCCat-), or SOD3 and catalase in combination (ASCdKO). Cells exposed to UW ± ASC-S at 4°C for 4 hours, were allowed to recover in standard iCM culture media for 8 hours prior to data collection. (B, C) Analysis of iCM beating velocity (B) and prevalence of cleaved caspase-3+ iCM (C). Cells exposed to UW ± ASC-S at 4°C for 4 hours were allowed to recover in standard iCM culture media (FM) for 24 hours prior to data collection and staining. (D, E) Dynamics of ROS (D) and H2O2
(E) accumulation in the cytoplasm of iCM exposed to UW alone or supplemented with either ASCC-S or ASCdKO-S (secretome produced by ASC silenced for SOD3 and catalase in combination) at 4°C for 4 hours, followed by recovery in FM. Time “0” represents the moment when UW solution was exchanged to FM. Statistics: (A-E): one-way ANOVA with Tukey’s post hoc test; n=6 for each graph; *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.